| 刘祥华,罗湘筠,李文倩.基于磷脂酰肌醇 3?激酶/蛋白激酶 B信号通路探讨针刺对超负荷运动致骨骼肌损伤大鼠氧化应激损伤及骨骼肌细胞凋亡的影响[J].安徽医药,2020,24(12):2443-2448. |
| 基于磷脂酰肌醇 3?激酶/蛋白激酶 B信号通路探讨针刺对超负荷运动致骨骼肌损伤大鼠氧化应激损伤及骨骼肌细胞凋亡的影响 |
| Effects of acupuncture on oxidative stress injury and apoptosis of skeletal muscle cells in rats with skeletal muscle injury caused by overload exercise based on phosphatidylinosital 3?kinase/protein kinase B signaling pathway |
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| DOI：10.3969/j.issn.1009?6469.2020.12.028 |
| 中文关键词: 运动损伤 肌，骨骼 针刺疗法 骨骼肌损伤 PI3K?Akt信号通路 氧化应激 磷脂酰肌醇 3?激酶 超氧化物歧化酶 琥珀酸脱氢酶 谷胱甘肽过氧化物酶 大鼠， Sprague?Dawley |
| 英文关键词: Athletic Injuries Muscle,skeletal Acupuncture therapy Skeletal muscle injury PI3K?Akt signaling pathway Oxidative stress PI3K Superoxide dismutase Succinic dehydrogenase Glutathione peroxidase Rats,Sprague?Dawley |
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| 中文摘要: |
| 目的探究针刺对超负荷运动致骨骼肌损伤大鼠氧化应激损伤及骨骼肌细胞凋亡的影响，初步了解针刺对超负荷运动致骨骼肌损伤的治疗机制。方法于 2018年 12月至 2019年 6月选取 60只 6周龄 SPF级 SD大鼠，采用随机数字表法将 SD大鼠分为四组:空白组(Control，C)、超负荷运动组(Overload，O)、针刺组(Acupuncture，A)和超负荷运动 +针刺组(Overload and Acupuncture，OA)，各 15只； O组及 OA组实行超负荷运动，运动结束立即针刺 A组及 OA组足三里、阳陵泉、内关和肾俞穴，留针 20 min，持续 14 d；检测各组 SD大鼠骨骼肌中超氧化物歧化酶(SOD)、丙二醛、琥珀酸脱氢酶(SDH)和谷胱甘肽过氧化物酶(GSH?Px)活性；使用流式细胞仪检测 SD大鼠骨骼肌细胞凋亡情况，使用蛋白质印迹法检测细胞凋亡相关蛋白 B细胞淋巴瘤 /白血病 ?2(Bcl?2)、 Bcl?2相关 X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶 3(Caspase?3)活性；检测 SD大鼠血清中肌酸激酶水平并使用苏木精 ?伊红(HE)染色观察 SD大鼠骨骼肌情况；使用蛋白质印迹法检测磷脂酰肌醇 3?激酶 /蛋白激酶 B(PI3K?Akt)信号通路相关蛋白表达情况。结果相比 C组丙二醛水平(4.75±0.29)mol/L、骨骼肌细胞凋亡水平(9.89±1.02)%和肌酸激酶(457.54± 45.02)kU/L，O组丙二醛水平(13.97±0.55)mol/L、骨骼肌细胞凋亡水平(32.58±3.50%)、 Bax、Caspase?3蛋白表达水、血清肌酸激酶水平(1985.50±88.87)kU/L均显著升高(P＜0.05)；相比 C组 GSH?Px水平(142.25±8.39)kU/L、SOD水平(2.50±0.25)kU/L、SDH水平(35.75±8.78)kU/L，O组 GSH?Px水平(55.68±6.97)kU/L、SOD水平(1.02±0.17)kU/L、SDH水平(10.58±6.50)kU/L、Bcl?2、 PI3K、磷酸化蛋白激酶 B(p?AKT)、哺乳动物雷帕霉素靶蛋白(m?TOR)水平均显著降低(P＜0.05)Akt蛋白表达差异无统计学意义(P＞0.05)；相比 O组， A组丙二醛水平(4.99±0.32)mol/L、骨骼肌细胞凋亡水平(11.52±1.02)%、B，ax、Caspase?3蛋白表达水、血清肌酸激酶水平均(557.56±54.25)kU/L显著降低， GSH?Px水平(130.25±7.14)kU/L、SOD水平(2.38±0.30)kU/L、SDH水平(31.25±5.50)kU/L、Bcl?2、PI3K、p?AKT、m?TOR蛋白水平显著升高(P＜0.05)Akt蛋白表达差异无统计学意义(P＞0.05)；相比 A组， OA组丙二醛水平(7.89±0.41)mol/L、骨骼肌细胞凋亡水平(22.58±2.51)%、B，ax、Caspase?3蛋白表达水、血清肌酸激酶水平(1402.51±64.50)kU/L均显著升高， GSH?Px水平(135.58±8.14)kU/L、SOD水平(2.10±0.21)kU/L、SDH水平(24.28±4.99)kU/L、 Bcl?2、PI3K、p?AKT、m?TOR蛋白水平显著降低(P＜0.05)，Akt蛋白表达差异无统计学意义(P＞0.05)；相比 O组， OA组丙二醛水平、骨骼肌细胞凋亡水平、 Bax、Caspase?3蛋白表达水、血清肌酸激酶平均显著降低， GSH?Px水平、 SOD水平、 SDH水平、 Bcl? 2、PI3K、p?AKT、m?TOR蛋白水平显著升高(P＜0.05)，Akt蛋白表达差异无统计学意义(P＞0.05)。 HE染色观察结果显示: O组纤维弯曲，肌红蛋白溶解，肌细胞核固缩、溶解，肌束膜破裂； OA组少量肌纤维断裂溶解弯曲，余未见异常。结论针刺可以通过调控 PI3K?Akt信号通路抑制 SD大鼠氧化应激和骨骼肌细胞凋亡来实现治疗超负荷运动致骨骼肌损伤。 |
| 英文摘要: |
| Objective To explore the effects of acupuncture on oxidative stress injury and apoptosis of skeletal muscle cells in ratswith skeletal muscle injury caused by overload exercise，and to preliminarily understand treatment mechanism of acupuncture for skeletal muscle injury caused by overload exercise.Methods Sixty 6?week?old SPF?level SD rats were enrolled from December 2018 and June 2019，and were assigned into four groups by random number table method:blank group(Control，C)，overload exer? cise group(Overload，O)，acupuncture group(Acupuncture，A)and overload exercise + acupuncture group(Overload and Acu? puncture，OA)，15 cases in each group.O group and OA group were given overload exercise.Immediately after end of exercise，acu? puncture was performed at Zusanli，Yanglingquan，Neiguan and Shenshu acupoints in group A and OA group，retaining needle for 20 min，lasting for 14 d.The activities of superoxide dismutase(SOD)，malondialdehyde(MDA)，succinate dehydrogenase(SDH) and glutathione peroxidase(GSH?Px)in skeletal muscle of each group were detected.Flow cytometry was applied to detect apopto?sis of skeletal muscle cells.Western blot was applied to detect activities of apoptosis?related proteins［B?cell lymphoma/leukemia?2(Bcl?2)，Bcl?2 related X protein(Bax)，cysteinyl aspartate specific protease3(Caspase?3)］.The level of serum creatine kinase was detected.The skeletal muscle conditions of rats were observed by hematoxylin?eosin(HE)staining.The expression of PI3K?Akt sig? naling pathway?related protein was detected by Western blot.Results Compared with MDA level［(4.75±0.29)mol/L］，apoptosislevel of skeletal muscle cells［(9.89±1.02)%］and creatine kinase［(457.54±45.02)kU/L］in group C，levels of MDA［(13.97±0.55)mol/L］，apoptosis level of skeletal muscle cells［(32.58±3.50%)］，expressions levels of Bax and Caspase?3 proteins，and lev? el of serum creatine kinase［(1985.50±88.87) kU/L］ in group O were significantly increased.Compared with GSH?Px level［(142.25±8.39)kU/L］，SOD level［(2.50±0.25)kU/L］and SDH level［(35.75±8.78)kU/L］in group C，GSH?Px level［(55.68±6.97)kU/L］，SOD level［(1.02±0.17)kU/L］，SDH level［(10.58±6.50)kU/L］，Bcl?2，phosphatidylinositol 3?kinase(PI3K)，phos? phorylated protein kinase B(p?AKT)and level of mammalian target of rapamycin(m?TOR)in group O were significantly de? creased(P＜0.05)，and there was no significant difference in expression of Akt protein between the two groups(P＞0.05).Com? pared with group O，MDA level［(4.99±0.32)mol/L］，apoptosis level of skeletal muscle cells［(11.52±1.02)%］，expression level of Bax and Caspase?3 proteins，and level of serum creatine kinase［(557.56±54.25)kU/L］in group A were significantly decreased， GSH?Px level［(130.25±7.14)kU/L］，SOD level［(2.38±0.30)kU/L］，SDH level［(31.25±5.50)kU/L］，Bcl?2，PI3K，p?AKT and m?TOR levels were significantly increased(P＜0.05)，and there was no significant difference in expression of Akt protein between the two groups(P＞0.05).Compared with group A，MDA level［(7.89±0.41) mol/L］， apoptosis level of skeletal muscle cells［(22.58±2.51)%］，expression levels of Bax and Caspase?3 proteins，and level of serum creatine kinase［(1402.51±64.50)kU/L］ in group OA were significantly increased，GSH?Px level［(135.58±8.14)kU/L］， SOD level［(2.10±0.21) kU/L］， SDH level［(24.28±4.99)kU/L］，Bcl?2，PI3K，p?AKT and m?TOR proteins were significantly decreased(P＜0.05)，and there was no signifi? cant difference in expression of Akt protein between the two groups(P＞0.05).Compared with group O，levels of MDA，skeletal muscle cell apoptosis，Bax and Caspase?3 protein，and level of serum creatine kinase in group OA were significantly decreased， while levels of GSH?Px，SOD，SDH，Bcl?2，PI3，p?AKT and m?TOR proteins were significantly increased(P＜0.05).There was no significant difference in expression of Akt protein between the two groups(P＞0.05).The observation results of HE staining showed that there were fiber waviness，myoglobin dissolution，muscle corpuscle pyknosis and dissolution，and perimysium rupturing in group O.In OA group，there were few muscle fibers dissolution，rupturing and waviness.And there were no abnormalities in the rest.Con? clusion Acupuncture can inhibit oxidative stress and skeletal muscle cell apoptosis in rats by regulating PI3K?Akt signaling path?way to treat skeletal muscle injury caused by overload exercise. |
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